IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ rothgiesser-2010-1 SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. in vitro In vitro deacetylase assay deacetylase assay western blot western blot Immuno blot Immuno blot Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Prior to the in vitro deacetylation assay, recombinant p65 was acetylated in vitro by incubation with recombinant human p300 (as described in PMID:11430825). Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch J. Cell. Sci. (0021-9533) 2010 Only protein-protein interactions imex curation rothgiesser-2010-2 SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. human-thp-1 Homo sapiens acute monocytic leukemia cells thp-1 Human acute monocytic leukemia cell line Human acute monocytic leukemia established from the peripheral blood of a 1-year-old boy with acute monocytic leukemia (AML) at relapse in 1978. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch J. Cell. Sci. (0021-9533) 2010 Only protein-protein interactions imex curation rothgiesser-2010-3 SIRT2 regulates NF-κB dependent gene expression through deacetylation of p65 Lys310. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy immunocytochemistry immunocytochemistry Rothgiesser K.M., Erener S., Waibel S., Lüscher B., Hottiger M.O. IBD - Inflammatory bowel disease The authors also show acetylation of p65 at lys-310 by p300 in an in vitro acetylation assay; however, the origin of the p300 construct used in this experiment could not be established, hence this interaction is not captured here. Mammalian expression vectors for human SIRT cDNA's are described in PMID:16079181, while those for human p65 and p65 mutants are described in PMID:18263619. HA-tagged SIRT2 is described in PMID:18332217. hottiger@vetbio.uzh.ch J. Cell. Sci. (0021-9533) 2010 Only protein-protein interactions imex curation sir2_human NAD-dependent protein deacetylase sirtuin-2 SIR2-like protein 2 SIRT2 SIR2L SIR2L2 Regulatory protein SIR2 homolog 2 protein protein human Homo sapiens Human MAEPDPSHPLETQAGKVQEAQDSDSDSEGGAAGGEADMDFLRNLFSQTLSLGSQKERLLDELTLEGVARYMQSERCRRVICLVGAGISTSAGIPDFRSPSTGLYDNLEKYHLPYPEAIFEISYFKKHPEPFFALAKELYPGQFKPTICHYFMRLLKDKGLLLRCYTQNIDTLERIAGLEQEDLVEAHGTFYTSHCVSASCRHEYPLSWMKEKIFSEVTPKCEDCQSLVKPDIVFFGESLPARFFSCMQSDFLKVDLLLVMGTSLQVQPFASLISKAPLSTPRLLINKEKAGQSDPFLGMIMGLGGGMDFDSKKAYRDVAWLGECDQGCLALAELLGWKKELEDLVRREHASIDAQSGAGVPNPSTSASPKKSPPPAKDEARTTEREKPQ tf65_human Transcription factor p65 Nuclear factor NF-kappa-B p65 subunit RELA NFKB3 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 protein protein human Homo sapiens Human MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS sir1_human NAD-dependent protein deacetylase sirtuin-1 NAD-dependent protein deacylase sirtuin-1 SIRT1 SIR2L1 SIR2-like protein 1 Regulatory protein SIR2 homolog 1 protein protein human Homo sapiens Human MADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLERSPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNGPGLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESDEEDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQIVINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYARLAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLLRNYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPADEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEVPQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRTQKELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCMEEKPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISRRLDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESEIEEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS q8ixj6-2 NAD-dependent protein deacetylase sirtuin-2 SIRT2 SIR2L SIR2L2 SIR2-like protein 2 Regulatory protein SIR2 homolog 2 protein protein human Homo sapiens Human MDFLRNLFSQTLSLGSQKERLLDELTLEGVARYMQSERCRRVICLVGAGISTSAGIPDFRSPSTGLYDNLEKYHLPYPEAIFEISYFKKHPEPFFALAKELYPGQFKPTICHYFMRLLKDKGLLLRCYTQNIDTLERIAGLEQEDLVEAHGTFYTSHCVSASCRHEYPLSWMKEKIFSEVTPKCEDCQSLVKPDIVFFGESLPARFFSCMQSDFLKVDLLLVMGTSLQVQPFASLISKAPLSTPRLLINKEKAGQSDPFLGMIMGLGGGMDFDSKKAYRDVAWLGECDQGCLALAELLGWKKELEDLVRREHASIDAQSGAGVPNPSTSASPKKSPPPAKDEARTTEREKPQ h4_human Histone H4 H4-16 H4C1 H4C11 H4C12 H4C13 H4C14 H4C15 H4C2 H4C3 H4C4 H4C5 H4C6 H4C8 H4C9 H4/A H4/B H4/C H4/D H4/E H4/G H4/H H4/I H4/J H4/K H4/M H4/N H4/O H4F2 H4FA H4FB H4FC H4FD H4FE H4FG H4FH H4FI H4FJ H4FK H4FM H4FN H4FO HIST1H4A HIST1H4B HIST1H4C HIST1H4D HIST1H4E HIST1H4F HIST1H4H HIST1H4I HIST1H4J HIST1H4K HIST1H4L HIST2H4 HIST2H4A HIST2H4B HIST4H4 protein protein human Homo sapiens Human MSGRGKGGKGLGKGGAKRHRKVLRDNIQGITKPAIRRLARRGGVKRISGLIYEETRGVLKVFLENVIRDAVTYTEHAKRKTVTAMDVVYALKRQGRTLYGFGG rela-sirt2 1765176 n/a 1765179 enzyme enzyme neutral component neutral component c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. n/a 1765180 enzyme target enzyme target substrate substrate neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false required acetyllys-310 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain false required acetyllys-314 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain false required acetyllys-315 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain false spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. deacetylation deacetylation reaction deacetylation false false false Fig. 2b, Suppl. Fig. S1a, S1e NAD+ (CHEBI:15846) SIRT2 deacetylated p65 in vitro at lysine-310, lysine-314 and lysine-315. Q8IXJ6 is required for deacetylation of Q04206 Q8IXJ6 decreases activity of Q04206 CHEBI:15846 increases activity of Q8IXJ6 P01375 increases binding of Q8IXJ6 to Q04206 rela-sirt1-4 1765176 n/a 1765181 enzyme enzyme neutral component neutral component c-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. n/a 1765180 enzyme target enzyme target substrate substrate neutral component neutral component required acetyllys-310 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain false required acetyllys-315 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false spofr-sf_21 Spodoptera frugiperda insect cells sf_21 Insect cells derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae. full name: IPLB-SF-21-AE cells are susceptible to Baculovirus infection. deacetylation deacetylation reaction deacetylation false false false Suppl. Fig. S1a, S1e SIRT1 deacetylated p65 in vitro at lysine-310 and lysine-315. While p65 was deacetylated in vitro at lysine-310 by SIRT2 after 30 minutes, deacetylation by SIRT1 required longer incubation time (120 minutes). NAD+ (CHEBI:15846) Q96EB6 is required for deacetylation of Q04206 rela-sirt2-1 1765177 n/a 1765180 unspecified role unspecified role bait bait n/a 1765179 unspecified role unspecified role prey prey physical association physical association false false false Fig. 3b tumor necrosis factor alpha (human) (TNF-alpha) (P01375) hist1h4a-sirt2 1765178 n/a 1765182 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false nuclear localisation signal tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q8IXJ6-2:p.[Leu4Ala;Leu12Ala;His150Tyr] mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n/a 1765183 unspecified role unspecified role neutral component neutral component possible acetyllys-17 naclys N-acetylated L-lysine NAcLys certain certain sequence position certain certain certain sequence position certain false colocalization colocalization false false false Suppl. Fig. S2a The Materials and Methods section states that the nuclear export signal (NES) of SIRT2 was disrupted by mutating K4 and K12 to alanines; however, the SIRT2 (isoform 2 with the NES at the N-terminus) contains leucine residues in these positions, which are a hallmark of NES motifs, hence these mutations are captured here as leucine-to-alanine mutations.